BACE1 enzyme activity detection was performed according to the manufacturer′s instructions (β-Secretase Activity Fluorometric Assay Kit, Sigma-Aldrich). Briefly, the lysis buffer of the primary cells was centrifuged at 10,000 g for 5 min, and a portion was used for the quantitative protein experiment with the BCA method. Then 50 μL of cell lysis was added, followed by the addition of reaction buffer and substrate. Fluorescence for each sample was measured using the full-wavelength microplate reader. The analyses were repeated at least twice in all samples.
β secretase activity fluorometric assay kit
Manufactured by Merck Group
The β-Secretase Activity Fluorometric Assay Kit is a laboratory tool used to measure the enzymatic activity of β-secretase, also known as BACE1. This kit utilizes a fluorogenic substrate to quantify the level of β-secretase activity in a sample. The assay provides a sensitive and reliable method for researchers to study the function and regulation of this important enzyme.
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Lab products found in correlation
4 protocols using β secretase activity fluorometric assay kit
1
Fluorometric Assay for BACE1 Activity
2
β-Secretase Activity Assay in NPC1 Mice
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β-secretase activity was assayed using a β-Secretase Activity Fluorometric Assay Kit (Sigma Aldrich). Cerebellar lobule samples were isolated from Npc1+/+ and Npc1−/− mice as described above, suspended in Extraction Buffer, and individual lobule lysates were prepared using a motor-driven pestle. For each lobule, four replicates of 10 µg lysate were loaded into a 96 well plate, and the assay was run according to the manufacturer’s instructions. The fluorometric results were obtained on a FLUOstar Omega (BMG Labtech, Ortenberg, Germany) with an excitation of 355 nm and an emission of 520 nm.
3
Quantifying Alzheimer's Biomarkers
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Aβ40 and Aβ42 levels were detected with commercial ELISA kits (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions. Activities of β-secretase and γ-secretase were measured with a β-secretase activity fluorometric assay kit (Sigma-Aldrich) and a γ-secretase activity assay kit (R&D Systems), respectively, following the manufacturers’ protocols.
4
BACE1 Activity Fluorometric Assay
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BACE1 enzyme activity detection was performed according the manufacturer's instructions (β-Secretase Activity Fluorometric Assay Kit, Sigma-Aldrich, MAK237). Briefly, cell lysis products were centrifuged at 10,000 g for 5 min, and a portion was used for protein quantitative experiment with BCA method. 50 μL of cell lysis was used, followed by the addition of reaction buffer and substrate. The fluorescence of each sample was detected using a full-wavelength microplate reader.
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